Immunostaining of Human Fetal Central Nervous Cells With the Anti-Human p75 pAb
Fix human fetal central nervous system (CNS) cells on coverslips for 10 minutes in 4%
paraformaldehyde.
Wash 2 times with PBS.
Permeabilize for 10 minutes in PBS + 0.1% Triton.
Wash 3 times with PBS.
Block for 30 minutes with PBS + 3% goat serum.
Incubate in Anti-Human p75 pAb diluted 1:50 in PBS + 3% goat serum for 60 minutes. For
a control, use normal Rabbit IgG
Wash 3 times with PBS.
Incubate in secondary antibody (Donkey anti-rabbit Cy3 Jackson Immunoresearch
Cat.#711-165-152) diluted 1:500 for 30 minutes. Do not expose slides to
light once the secondary antibody has been applied.
In the dark, wash 3 times with PBS.
Mount with Vectashield Antifade + DAPI (Vector Labs)
